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1 Electronic upplementary Material (EI) for rganic & Biomolecular Chemistry. This journal is The Royal ociety of Chemistry 2014 upplemental Information olid-phase ynthesis of Peptoid-like ligomers Containing Diverse Diketopiperazine Units ujit uwal and Thomas Kodadek* Materials and Instruments: Amino acid esters were purchased from either from Aldrich, Acros, Advanced chemtech and Astatech. () and (R)-2-bromo-propionic acid were purchased from Aldrich. Mono- -tboc-ethylenediamine was bought from Astatech Inc. Primay amines were purchased from Advanced Chemtech, Aldrich, Acros, TCI. Rest of the other chemicals and reagents were purchased either from Aldrich or Fisher cientific. Rink amide MBA resin (0.5 mmol/g) and Tentagel macrobead- 2 (160 µg, 0.4 mmol/g) resin were obtained from Chemprep and rapp-polymere, respectively. Disposable fritted columns (5 ml, 50 ml) for peptoid syntheses were bought from Intavis AG. Microwave assisted peptoid syntheses were accomplished utilizing 10% of 1550 W household microwave (GE model JE 1860 B04). PLC purification of crude peptoids after acid cleavage from Rink amide resin was carried out in Water 1525 binary PLC pumps and a 2487 dual absorbance detector, or a 2998 photodiode array detector. Buffer A ( 2 with 5% C 3 C and 0.1% trifluoroacetic acid (TFA) and buffer B (5m, 250 x 4.6 mm, Alltech) and Apollo C-18 5µ preparative column. Purities of the compounds were assessed by analytical PLC under UV (214), otherwise mentioned in description of the spectrum. M and M/M were recorded in MALDI -TF (4800 Proteomics Analyser, Applied Biosystems) or EI-M (Waters). In M/M, the signal-to-noise ratio threshold set to 10. Cyano-4-hydroxycinnamic acid (CCA) or charcoal was used as MALDI matrices (Aldrich). 1 and 13 C MR spectra were recorded in Bruker 400 instrument operating at 400 Mz for 1 and 100 Mz for 13 C MR using Deuterated-methanol (CD 3 D) or Deuterated-Dimethylsulfoxide (DM) (Cambridge) as a solvent. ynthesis of 2, 5-DKP (4 a-f) cheme 1 Br o C, 2 2. TFA:DCM:TI (60:38:2) Amino acid ester DMF, 37 o C, R R 4 a-f R' 1. BAA, DIC 37 o C, 2x 30 min 2. Primary amine DMF, 37 o C, 1 h R" Where, R"= C 3 or C 2 5 a. R = ()-Me, R' = Benzyl R b. R = ()-4-hydroxybenzyl, R'= 2- methoxy ethylamine Ph e. R = ()-Me, R' = c. R = ()-Me, R' = d. R = (R)-Me, R' = Ph f. R = (R)-Me, R' = R' R" 1

2 2 4a Rink amide MBA resin (1 g, 0.7 mmol) was swelled in DMF. Fmoc group was deprotected with 20% piperidine in DMF (2X, 15 mins). The resultant primary amine was coupled with bromoacetic acid (BAA, 2M) activated with diisopropylcarbodiimide (DIC, 2 M) in, -dimethylformamide (DMF). ubsequently, the bromide was displaced with desired Cl salt of amine acid ester in DMF (2 M) along with DIEA (1.5X with respect to ester). The secondary amine was further coupled with bromoacetic acid (2M), activated with DIC (3M), 37 C (2X, 15 min). The bromide was displaced with a solution of desired primary amine (viz benzyl amine in this case) in DMF at 37 o C, 1 h. At this stage, the reaction mixture further incubated at 50 o C overnight so that the secondary amine thus generated after 2 reaction undergo addition elimination at the ester thus generating six-membered 2,5 diketopiperazine. Finally, the solid support was washed with DMF, DCM and ethanol, and subjected to cleave with 50% TFA in DCM along with 2% triisopropylsilane. A crude DKP obtained after the solvent was evaporated under Argon and ether precipitation. A crude dried compound finally was subjected to MR study without further PLC purification by dissolving in d-methanol. 1 MR (400 MZ, D 6 -DM) δ (m, 5), 4.7 (d, 1, J=14.7), 4.5 (d, 1, J=14.7), 4.3 (d, 1, J=16.7), 4.12 (q, 1), 4.0 (d, 1, J=17.7), 3.9 (d, 1, J=16.7), 3.82 (d, 1, J= 17.5), 1.5 (d, 3, Me, J=7.2); 13 CMR (100 Mz, DM) δ 172.3, 169.2, 166.5, 136.8, 129.9, 59.19, 50.3, 49.7, 47.8, 18.1; MALDI-TF M (bs.=ma + ) ( expected). Figure 1: 1 MR of compound 4a 2

3 Figure 2: CY of 4a Figure 3: 13 CMR of 4a Figure 4: Dept of 4a 3

4 2 Exact Mass: Fig 5: MALDI-M of compound 4a 2 4b pectral data for compound 4b: A crude dried compound was dissolved in in d-dm. 1 MR (400 MZ, D 6 -DM) δ 6.83 (d, 2, J=8.3), 6.66 (d, 2, J=8.3) 4.35 (d, 1, J=16.4), 4.15 (t, 1), 3.6 (d, 1, J=16.4), 3.32 (m, 1, J=17.1), 3.5 (d, 1), 3.35 (m, 1), 3.3 (m, 1), 3.2 (s, 3), 3.15 (m, 1), 3.0 (dd, 1, J=14), 2.9 (dd, 1, J=14), 2.6 (d, 1, J=17.1); 13 CMR (100 Mz, DM) δ 169.2, 165.3, 164.0, 156.5, 130.7, 124.9, 115.0, 69.2, 61.7, 57.8, 49.5, 45.8, 44.9, 35.5; EI-M(bs. Ma + ) = (Expected= ). Figure 6: 1 MR of 4b 4

5 Figure 7: 13 CMR of compound 4b Figure 8: 13 C-DEPT of compound 4b Figure 9: CY of compound 4b 5

6 2 Exact Mass: Fig 10: MALDI-M of compound 4b pectral data for compound 4c and 4d: For compound 4d: A PLC purified compound was dissolved in in d-chloroform. 1 MR (400 MZ) δ (m, 5), (m, 2), 3.90 (d, 1), 3.60 (d, 1), 3.35 (d, 1), 3.23 (m, 1), 3.02 (m, 1), 1.45 (d, 3), 1.29 (d, 3); 13 CMR (100 Mz, DM) δ 172.6,169.6, 166.9, 63.1, 59.2, 52.3, 47.7, 45.5, 17.7, 13.4; MALDI-TF M (bs. Ma + ) (Expected = expected). For compound 4c: A PLC purified compound was dissolved in in d-chloroform. 1 MR (400 MZ) δ (m, 5), 4.0 (d, 1), (m, 3), 3.83 (d, 1), 3.61 (d, 1), (m, 1), (m, 1), 1.3 (d, 3), 1.2 (d, 3); MALDI-TF M (bs. Ma + ) (Expected = expected). 6

7 c Me 2 4d b Me a Me 2 4c b Me Figure 11: 1 MR of compound 4c and 4d c Me 2 4d b Me a Me 2 4c b Me Figure 12: 13 CMR of compound 4c and 4d 7

8 2 4d c Me b Me a Me 2 b Me 4c Figure 13: 13 CMR of compound 4c and 4d 2 4c a Me b Me 2 4c a Me b Me Figure 14: 13 CMR of compound 4c and 4d 8

9 2 4d c Me b Me a Me 2 4c b Me Figure 15: PLC and EI-M of compound 4c and 4d 2 4e pectral data for compound 4e: A crude dried compound was dissolved in in d4-methanol. 1 MR (400 MZ, D 4- Me) 4.6 (m, 1), 4.3 (d, 1), 4.14 (d, 1), 4.06 (q, 1), 4.02 (d, 1), 3.9 (d, 1), 3.64 (m, 1), 1.49 (d, 1), 1.18 (d, 1); 13 CMR (100 Mz, DM) δ 172.6,169.6, 166.9, 63.1, 59.2, 52.3, 47.7, 45.5, 17.7, 13.4; MALDI-TF M (bs. Ma + ) (Expected = ). Figure 16: 1 MR of compound 4e 9

10 Figure 17: Cosy of compound 4e Figure 18: 13 CMR of compound 4e Fig 19: MALDI-M of compound 4e 10

11 2 4f pectral data for compound 4f: A crude dried compound was dissolved in D6-DM. 1 MR (400 MZ, D 6 -DM) 4.6 (m, 1), 4.3 (d, 1), 4.14 (d, 1), 4.06 (q, 1), 4.02 (d, 1), 3.9 (d, 1), 3.64 (m, 1), 1.49 (d, 1), 1.18 (d, 1); 13 CMR (100 Mz, DM) δ 169.3, 166.8, 164.2, 61.6, 59.7, 43.8, 40.0, 45.5, 17.2, 13.1; MALDI-TF M (bs. Ma + ) ( expected). Figure 20: 1 MR of compound 4f 11

12 Figure 21: CY of compound 4f Figure 22: 13 C MR of compound 4f 12

13 Figure 23: DEPT of compound 4f Figure 24: MALDI-TF M of compound 4f Figure 25: PLC of compound 4f 13

14 olid phase synthesis protocol for 2,5-DKP containing analog utilized for purity test: cheme BAA, DIC, DMF, 37 o C,.. 2. Amino acid ester, DMF, o C Cl R' 1. BAA, DIC, DMF 30 min, 37 o C 2 Cl o C, TFA:DCM:TI 7 (60:38:2), 2 h. R' Cl Cl Cl Cl R' = Ph,,,,, a b c d e,,, f g h i 2 Rink amide MBA resin (100 mg, 0.07 mmol) was swelled in DMF. After the fmoc group deprotection with 20% piperidine in DMF (2X, 15 mins), a dimer peptoid was synthesized by conventional peptoid synthesis protocol. nto the resultant dimer additional bromoacetic acid was added and bromide was displaced by amino acid ester. The secondary amine was further coupled with DIC activated bromoacetic acid at 37 C (2X, 15 min). The bromide was displaced primary amine (2 M) in DMF at 37 o C, 1 h. To afford DKP, the reaction mixture was further incubated at 50 o C overnight. Completion of the reaction was confirmed by Chloranil test. The compound thus obtained was further attempted for chain extension with DIC activated BAA followed by displacement with primary amine. At this stage, the bead was acid-cleaved and subjected for PLC to test the purity of DKP formation. PLC fractions were subjected for MALDI-TF analysis to confirm the presence of desired molecule. DKP-Gly: Figure 26: PLC of 7a showing retention time at 5 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. 14

15 DKP-Ala: Figure 27: PLC of 7b showing retention time at 5 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. DKP-Ileu: Figure 28: PLC of 7c showing retention time at 5 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. DKP-Leu: Figure 29: PLC of 7d showing retention time at 5 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. 15

16 DKP-PhGly: Figure 30: PLC of 7e showing retention time at 35 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. DKP-Tyr: Figure 31: PLC of 7f showing retention time at 5 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. DKP-is: Figure 32: PLC of 7g showing retention time at 26 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. 16

17 DKP-Trp Figure 33: PLC of 7h showing retention time at 35 min. with MALDI-TF (in the middle) showing the molecular mass after the purification DKP-Lys Figure 34: PLC of 7i showing retention time at 26 min. with MALDI-TF (in the middle) showing the molecular mass after the purification. PLC and MALD-TF with M/M data of few compounds that are used to test the feasibility and purity for chain extension purpose. In these particular examples alloc group was utilized as a protecting group instead of tboc- group. The allocgroup was deprotected as explained in our previous paper (rg. Biomol. Chem., 2013, 11, ) / / / 729 Exact Mass: / 167 Figure 35: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 17

18 167/ / / / / / 905 Exact Mass: Figure 36: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound / / / 201 Cl Cl 520 / / 254 Exact Mass: Figure 37: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 18

19 947 / / / / / 662 Exact Mass: Figure 38: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. PLC and MALD-TF with M/M data of few compounds that resembles DKPhybrid compound library. 254 / / 388 CF 3 CF 3 2 CF 3 Ph 368 / 626 Exact Mass: Figure 39: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound 19

20 484 / / 693 CF / / CF 3 2 Exact Mass: Figure 40: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 610/ 384 CF 3 252/ / CF 3 Exact Mass: Figure 41: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. Protocol for peptoid library construction: DKP-Backbone library: Tentagel MB 2 (0.5 g, initial loading 0.4 mmol/g) was soaked in DMF for 30 min, RT. The beads were coupled with fmoc-methionine (5X) activated with,,, - Tetramethyl--(1- benzotriazol-1-yl)uronium hexafluorophosphate (BTU) (5X), - ydroxybenzotriazole (Bt) (8X) and Diisopropyl ethylamine (DIEA) (10x) in DMF. Mixture was incubated 3 h at room temperature. Fmoc-group was deprotected using 20% piperidine in DMF. Resultant primary amine was treated with BAA/DIC and the bromide was displaced with methoxyethylamine under microwave condition. Likewise, methylamine was added as additional two monomers to represent a linker region. The 20

21 main chain 2,5-DKP containing peptoid oligomer library (~ 40 K diversity) was created utilizing 12 different commercially available primary amines along with 8 different 2,5- DKP moieties. As a first unit of peptoid library, BAA/DIC was coupled to the secondary amine from the linker. The solid support was split into 12 different fractions after the beads were washed with DMF (5X) and DCM (5X). Each portion of the beads was separately treated with different primary amines (2M) to displace the bromide under microwave condition. The beads were washed as explained above. Resultant secondary amines were pooled and coupled with BAA/DIC. The beads were washed and split into 8 different vessels and treated them separately with amino acid esters. After additional round of BAA/DIC coupling, tboc-protected ethylenediamine was used to displace the bromide. The reaction mixture was incubated o C, 24h to afford 2,5-DKP formation. T-boc group was deprotected by 50% TFA in DCM. The resultant primary amine was separated into two halves and coupled with DIC-activated BAA or ()-2-bromopropionic acid separately. The bromides were pooled and split into 12 different fractions and displaced individually by primary amines (R ). Finally a peptoid oligomer library with 2,5-DKP in the main-chain was finally completed by final round to BAA/DIC coupling and bromide displacement with different sets of 12 primary amines (R, cheme ch2). 2 A R : R A R' R 2 R" R" CF F Ph CF Ph DKP-ide-chain library: The linker region and the first diversity unit in DKP-side-chain library were created as in the case of DKP-backbone library. After the first diversity element, the chain extension was carried out coupling with BAA/DIC followed by mono--tboc-ethylenediamine, as a later point of chain-extension as a side-chain. The secondary amine on the other hand was coupled with BAA and the solid support was split into 5 different reaction vessels. Each vessel was separately treated with amino acid ester. Beads were pooled and split into 8 21

22 fractions and treated individually with 8 different primary amines. At this stage the reaction mixture was subjected for 2,5-DKP ring formation under basic thermal condition as explained above. nce the cyclization was completed, the t-boc group was deprotected utilizing 50% TFA in DCM. Two additional peptoid monomers were added from the resultant primary amine utilizing conventional peptoid synthesis method via split and pool method. A R R A R R' R : 2 R" CF 3 2 Cl 2 2 Cl Ph 2 2 CF 3 2 Cl 2 Cl F DKP-terminated hybrid peptidomimetic library: The linker region and the first diversity unit in DKP-side-chain library were created as in the case of DKP-backbone library. The solid support was divided into 16 different vessels. In order to incorporate variety of diversity element at the second diversity element, following processes were executed, 1. ut of 16, two of the fractions were separately coupled with DIC activated ()-2- bromopropionic acid and separately treated with either mono--tboc-piperazine or ()-4- -Boc-2-methylpiperazine at 50 o C,.. The t-boc group was deprotected using 50% TFA in DCM prior to third diversity element was incorporated. 2. ne portion out of remaining fractions was coupled with BAA/DIC and bromide was displaced with -mono-t-boc-ethylenediamine. The secondary amine was coupled with DIC activated ()-2-bromo-3-phenyl-propionic acid. t-boc group was acid-deprotected and the reaction mixture was incubated under 10% DIEA overnight to afford cyclization to afford 2-oxopiperazine unit. 3. Remaining portion of the beads were divided into two fractions after pooling. Each 22

23 fraction was equilibrated in DCM and treated directly either with 3-chloromethyl benzoyl chloride (10X) or with 4-bromomethyl phenylsulfonyl chloride in DCM (5 ml). A solution of triethylamine (12X) in DCM (5 ml) was slowly added in the acid chloride solution at 0 o C. The reaction mixture was brought into the room temperature and allowed to stand for 3 hr. Each vessel was separately washed with DCM several times, pooled together and split into 13 different fractions. The halides were incubated with 13 different amines (R ) separately at 37 o C, 3 hr. After completion of above explained syntheses, all 16 vessels were pooled and coupled with BAA/DIC. The beads were further split into 13 portion and treated with 13 primary amines separately (R ), 37 o C, 1 h. Finally, library synthesis was accomplished by incorporating 2,5-DKP as a capping group. The formation of DKP was performed as explained above. At the terminal position, 56 different 2,5-DKPs as capping groups were generated with 7 different amino acid esters and 8 different primary amines. 23

24 DKP-hybrid library a. Linker R' b. Linker R' Linker Linker = 2. ()Me-Piperazine, DMF R' c. d. e. 2. Piperazine, DMF Linker Linker Linker R' R' a. b. c. d. e. 1. ()BPA, DIC, 1. ()BPA, DIC, DMF DMF Cl Cl Cl DIEA, DCM, 0 o C 2. R'- 2, DMF 50 o C, 2 h R' R' R' Ph DIEA, DCM, 0 o C 2. R'- 2, DMF 37 o C, 30 min 1. BAA, DIC, DMF 2. R' 2, DMF 3. DKP formation as shown in cheme 1 DKP Br DKP: R Linker R" R= ide-chain from alanine, leucine, lysine, phenylglycine, phenylalanine, histidine, tryptophan R' 1. BAA, DIC, DMF, 37 o C, 15 min 2. -tboc-ethylenediamine DMF, 37 o C, 30 min 3. Ph Br DMF, 37 o C, 2 h 4. 50% TFA in DCM, RT, 30 min 5. 10% DIEA in DMF 2 hr, 37 o C A R' 24

25 F 3 C R' Cl F 3 C R" CF Cl Cl 2 DKP: R R" R= ide-chain from alanine, leucine, lysine, phenylglycine, phenylalanine, histidine, tryptophan Cl Following are the structures of peptoids from DKP-backbone library, identifed after CBr mediated cleavage from the resins and subsequent MALDI M and M/M analyses. Each figure has a peptoid structure showing the mass of corresponding y and b fragments at particular substituent observed during Collision-Induced Dissociation (CID) of the parent peak: 359 / / / / 206 Exact Mass: Figure 42: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound 25

26 359 / / / / 178 Exact Mass: Figure 43: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound Cl Cl 359 / / / / / 230 Exact Mass: Figure 44: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 26

27 754 / / 795 CF / / 653 Exact Mass: Figure 45: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. F 359 / / / / 595 Exact Mass: Figure 46: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 27

28 329/ 737 Ph 855 / / 464 Exact Mass: Figure 47: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 359 / / 381 F 3 C 1053 / / 707 Exact Mass: Figure 48: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 28

29 751 / 389 Ph 974 / / 628 Exact Mass: Figure 49: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 359 / / / 395 CF / / 672 CF 3 Exact Mass: Figure 50: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 29

30 359 / / / / / Exact Mass: Figure 51: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound CF / / / / 175 Exact Mass: Figure 52: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. \ 30

31 1074 / / / 933 Exact Mass: Figure 53: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound 1056 / / / 933 Exact Mass: Figure 54: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 31

32 804 / / / / / 594 Exact Mass: Figure 55: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 359 / / / / / 627 Exact Mass: F Figure 56: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 32

33 2 358 / / 1079 Exact Mass: / / 166 Figure 57: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. DKP-side chain library: Ph 1210 / / / / / / Exact Mass: Figure 58: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 33

34 Ph 1230 / / / / 1119 Cl 588 / / 319 Exact Mass: Cl Figure 59: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound / / / / / / 320 Exact Mass: Figure 60: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 34

35 999 / / / / / / / 319 Exact Mass: Figure 61: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. F 3 C 1063 / / / / / / 319 Exact Mass: Figure 62: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 35

36 359 / / / / / 787 F 992 / 319 Exact Mass: Figure 63: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound / / / / / / 344 Exact Mass: Figure 64: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 36

37 1093 / / / / / 592 Exact Mass: / Figure 65: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. Cl Cl 1256 / / / / / 729 Exact Mass: / Figure 66: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 37

38 919 / / / / / / 237 Exact Mass: Figure 67: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 967 / / / / / 538 Exact Mass: Figure 68: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 38

39 ybrid DKP-capped library: 359 / / / / 546 Exact Mass: Figure 69: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 789 / / / / / 630 CF / 658 Exact Mass: Figure 70: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 39

40 2 712 / / / / / / 672 Exact Mass: Figure 71: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. CF / / / / / / 679 Exact Mass: CF3 Figure 72: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 40

41 2 456 / / / / / 464 Exact Mass: / 312 Figure 73: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 359 / / / / / 282 Exact Mass: Figure 74: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 41

42 372 / / / / 348 F 3 C 879 / 588 Exact Mass: Figure 75: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 524 / / / 781 Exact Mass: / 430 Figure 76: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 42

43 2 496 / / / / / 724 Exact Mass: / / 464 Figure 77: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 359 / / / / / 595 Exact Mass: / 623 Figure 78: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 43

44 763 / / / / / CF 3 Exact Mass: Figure 79: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 472 / / / / / 417 Exact Mass: / 320 Figure 80: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 44

45 2 381 / 1003 Ph 310 / / / / Exact Mass: ( + ) (a + ) Figure 81: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound 359 / / 546 Ph 686 / / 393 CF 3 Exact Mass: Figure 82: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 45

46 Ph Exact Mass: Figure 83: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. F 3 C 310 / / 883 Ph 826 / 437 Exact Mass: / 225 Figure 84: Top: MALDI-M data showing parent peak (left) of peptoid (right). Bottom: MALDI-TF M/M of the compound. 46

47 47